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    Congenitally dysplastic inferior rectus muscle.
    ( 2010-01-01) Kekunnaya, Ramesh ; Bansal, Rasheena ; Vemuganti, Geeta K.
    The authors report an unusual presentation of an idiopathic congenitally dysplastic inferior rectus muscle that responded well to surgical correction. Isolated unilateral enlargement of extraocular muscles is rare in children, and there is no definite logical explanation for its cause. A 20-month-old child presented with a congenitally enlarged posterior part of the right inferior rectus muscle with prominent hypotropia and enophthalmos since 10 months of age. Systemic disease work-up, ultrasound B-scan, computed tomography of the orbit and brain, and inferior rectus muscle biopsy were performed. Preoperatively, the child had severe hypotropia of the right eye with retraction of the globe. Work-up for systemic diseases was negative. Computed tomography scan showed thickening of the posterior two-thirds of the inferior rectus muscle. Muscle biopsy showed non-specific fibrotic changes. Strabismus surgery was undertaken at 2 years of age. Hypotropia was reduced significantly postoperatively. Compensatory head position was eliminated. Copyright 2010, SLACK Incorporated.
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    Evisceration in unsuspected intraocular tumors
    ( 2010-03-01) Rath, Suryasnata ; Honavar, Santosh G. ; Naik, Milind N. ; Gupta, Roshmi ; Reddy, Vijay A. ; Vemuganti, Geeta K.
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    Unstimulated diagnostic marrow tap - a minimally invasive and reliable source for mesenchymal stem cells
    ( 2010-03-01) Koppula, Purushotham Reddy ; Polisetti, Naresh ; Vemuganti, Geeta K.
    BMMNCs (bone marrow mononuclear cells) were isolated by density gradient centrifugation from unstimulated diagnostic marrow tap to propagate and characterize hBMSCs (human bone marrow stromal cells) and to explore their plasticity towards neuronal and other lineages. hBMSCs were characterized by flow cytometry for established markers, serially passaged and differentiated into adipo, osteo, chondro and neuronal lineages. Neural differentiation was analysed by RT-PCR (reverse transcriptase-PCR), ICC (immunocytochemistry) and Western blotting. The hBMSCs (n539) were spindle-shaped and immunoreactive for mesenchymal markers such as CD71, CD106, CD105, CD90 and Vimentin and negative for haematopoietic markers such as CD11c, CD34 and CD45. These cells showed differentiation into adipocytes, osteocytes and chondrocytes. Upon neuronal differentiation, hBMSCs expressed neuronal markers, i.e. b-III tubulin, GAP43 (growth-associated proteins), neurofilament by ICC, RT-PCR and Western blotting. Our study demonstrates that minimal volumes of unstimulated diagnostic marrow tap forms a minimally invasive and reliable source for isolation of BMMNCs to establish cultures of mesenchymal stem cells and expand them. The plasticity observed in these cells towards mesenchymal (adipogenic, osteogenic and chondrocytic) and non-mesenchymal lineage (neural) substantiates the nature of mesenchymal stem cells and warrants further studies to evaluate their functional role. © The Author(s) Journal compilation © 2010 Portland Press Ltd.
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    Isolation, characterization and differentiation potential of rat bone marrow stromal cells
    ( 2010-04-01) Polisetti, Naresh ; Chaitanya, V. G. ; Babu, Phanithi Prakash ; Vemuganti, Geeta K.
    Background: Bone marrow mesenchymal cells have been identified as a source of pluripotent stem cells with varying degrees of plasticity in humans. However, there are a few reports on rat-derived cells, which could be good models for the research purpose. We describe here a simple method of establishing the rat bone marrow stromal cells by the principle of adhesion and document their phenotype along with their differentiation potential to other lineages. Materials and Methods: Rat bone marrow stromal cells were isolated by three methods: direct plastic adherence, ficoll hypaque separation and a combination of both. The stromal cells obtained by these methods were characterized by fluorescent activating cell sorting (FACS) for established hematopoietic and non-hematopoietic markers. The cells obtained by combination method (combination of ficoll density gradient centrifugation and plastic adherence) were cultured and serially passaged. Transcriptional confirmation was done by reverse transcription polymerase chain reaction (RT-PCR) for vimentin and collagen type 1 alpha 1. Attempts were made to differentiate the marrow stromal cells into adipocytes, osteocytes and neuronal like cells. Results: Bone marrow samples from 10 rats yielded 4-5 million bone marrow mononuclear cells /ml per femur. Of the three methods tested, a combination method yielded good growth of spindle cells. The cells obtained by combined method showed high percentage of positivity for vimentin, fibronectin and CD90 and negative for hematopoietic markers. Further, RT-PCR confirmed vimentin and collagen type - 1 alpha 1 expression. Oil red O staining and Alizarin red staining confirmed adipocytic and osteogenic differentiation. On immunocytochemical analysis, the cells expressed nestin, -tubulin III, neurofilament and synaptophysin. Conclusion: Adequate quantities of rat marrow stromal cell cultures can be established by a simple method based on adhesion properties. Their phenotypic characteristics and plasticity support the evidence that they are mesenchymal stem cells with a distinct tendency for neural lineage.
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    Reply
    ( 2010-06-01) Kesarwani, Siddharth ; Murthy, Ramesh ; Vemuganti, Geeta